GSM7119490: PCW11 [scRNAseq]; Homo sapiens; RNA-Seq
Fetal eyes at 9–26 weeks of gestation were dissected from fetal and embryonic terminations, collected in DMEM/F-12 (Gibco, Australia), and transferred into a sterile Petri dish. The fetal eye tissue was then sectioned into pieces, followed by digestion in 3 mL of 2 mg/mL papain (brands) for approximately 20 min at 37°C. Using a 30 mL MACS Smart Strainer (Miltenyi), the dissociated fetal eyeball tissue was filtered to guarantee a single-cell suspension. Enzymatic digestion was stopped by the addition of an equal volume of 10% fetal bovine serum (Gibco, Australia) to DMEM/F-12 (Gibco, Australia). The cells were then pelleted and resuspended in an appropriate amount of PBS. A total of 74,571 cells were collected for scRNA-seq and 75,197 cells for scATAC-seq. Cells were loaded onto the 10X Chromium Single Cell Platform (10X Genomics) at a concentration of 1,000 cells per μl (Single Cell 3′ library and Gel Bead Kit v.3) as described in the manufacturer's protocol. Generation of gel beads in emulsion (GEMs), barcoding, GEM-RT clean-up, complementary DNA amplification and library construction were all performed as per the manufacturer's protocol. Qubit was used for library quantification before pooling. The final library pool was sequenced on the Illumina NovaSeq 6000 instrument using 150-base-pair paired-end reads.